Can we improve heterosis for root growth of maize by selecting parental inbred lines with different temperature behaviour?

Tolerance to high and low temperature is an important breeding aim for Central and Northern Europe, where temperature fluctuations are predicted to increase. However, the extent to which genotypes differ in their response to the whole range of possible temperatures is not well understood. We tested the hypothesis that the combination of maize (Zea mays L.) inbred lines with differing temperature optima for root growth would lead to superior hybrids. This hypothesis is based on the concept of 'marginal overdominance' in which the hybrid expresses higher relative fitness than its parents, summed over all situations. The elongation rates of axile and lateral roots of the reciprocal cross between two flint and two dent inbred lines were assessed at temperatures between 15°C and 40°C. Indeed, the cross between UH005 and UH250 with lateral root growth temperature optima at 34°C and 28°C, respectively, resulted in intermediate hybrids. At temperatures below and above 31°C, the hybrids' root growth was comparable to the better parent, respectively, thereby increasing temperature tolerance of the hybrid compared with its parents. The implications of and reasons for this heterosis effect are discussed in the context of breeding for abiotic stress tolerance and of putatively underlying molecular mechanisms. This finding paves the way for more detailed investigations of this phenomenon in future studies.

QTL involved in the partial restoration of male fertility of C-type cytoplasmic male sterility in maize.

Partial restoration of male fertility limits the use of C-type cytoplasmic male sterility (C-CMS) for the production of hybrid seeds in maize. Nevertheless, the genetic basis of the trait is still unknown. Therefore, the aim to this study was to identify genomic regions that govern partial restoration by means of a QTL analysis carried out in an F(2) population (n = 180). This population was derived from the Corn Belt inbred lines B37C and K55. F(2)BC(1) progenies were phenotyped at three locations in Switzerland. Male fertility was rated according to the quality and number of anthers as well as the anthesis-silking interval. A weak effect of environment on the expression of partial restoration was reflected by high heritabilities of all fertility-related traits. Partial restoration was inherited like an oligogenic trait. Three major QTL regions were found consistently across environments in the chromosomal bins 2.09, 3.06 and 7.03. Therefore, a marker-assisted counter-selection of partial restoration is promising. Minor QTL regions were found on chromosomes 3, 4, 5, 6 and 8. A combination of partial restorer alleles at different QTL can lead to full restoration of fertility. The maternal parent was clearly involved in the partial restoration, because the restorer alleles at QTL in bins 2.09, 6.04 and 7.03 originated from B37. The three major QTL regions collocated with other restorer genes of maize, a phenomenon, which seems to be typical for restorer genes. Therefore, a study of the clusters of restorer genes in maize could lead to a better understanding of their evolution and function. In this respect, the long arm of chromosome 2 is particularly interesting, because it harbors restorer genes for the three major CMS systems (C, T and S) of maize.

Root growth and nitrate-nitrogen leaching of catch crops following spring wheat.

Growing nitrogen (N) catch crops can reduce NO(3)-N leaching after cultivating cereals. The objective of this study was to relate NO(3)-N leaching to variation in the uptake of N and the size and distribution of the root systems of different catch crops species. In a 3-yr lysimeter experiment, phacelia (Phacelia tanacetifolia Benth.), sunflower (Helianthus annuus L.), and a Brassica species (yellow mustard [Brassica alba L.] or a hybrid of turnip rape [B. rapa L. spp. oleifera (DC.) Metzg.] and Chinese cabbage [B. rapa L. ssp. chinensis (L.) Hanelt]) were grown after the harvest of spring wheat under two levels of N supply. Bare soil lysimeters served as the control. Water percolation from the lysimeters and the NO(3)(-) concentration in the leachate were measured weekly from the sowing until the presumed frost-kill of the catch crops. Minirhizotrons were used to assess the spatial and temporal patterns of root growth from 0.10 to 1.00 m. The catch crop species differed in their shoot biomass, N uptake, total NO(3)-N leaching, and root growth. The results suggested that there was no strict relationship between the total NO(3)-N leaching of each catch crop species and the N uptake or parameters that indicate static characteristics of the root system. In contrast, the ranking of each catch crop species by parameters that indicate early root growth was inversely related to the ranking of each catch crop species in NO(3)-N leaching. The rapid establishment of the root system is essential for a catch crop following spring wheat to reduce the amount of NO(3)-N leaching after the harvest of spring wheat.

QTLs for the elongation of axile and lateral roots of maize in response to low water potential.

Changes in root architecture and the maintenance of root growth in drying soil are key traits for the adaptation of maize (Zea mays L.) to drought environments. The goal of this study was to map quantitative trait loci (QTLs) for root growth and its response to dehydration in a population of 208 recombinant inbred lines from the International Maize and Wheat Improvement Center (CIMMYT). The parents, Ac7643 and Ac7729/TZSRW, are known to be drought-tolerant and drought-sensitive, respectively. Roots were grown in pouches under well-watered conditions or at low water potential induced by the osmolyte polyethylene glycol (PEG 8000). Axile root length (L (Ax)) increased linearly, while lateral root length (L (Lat)) increased exponentially over time. Thirteen QTLs were identified for six seedling traits: elongation rates of axile roots (ER(Ax)), the rate constant of lateral root elongation (k (Lat)), the final respective lengths (L (Ax) and L (Lat)), and the ratios k (Lat)/ER(Ax) and L (Lat)/L (Ax.) While QTLs for lateral root traits were constitutively expressed, most QTLs for axile root traits responded to water stress. For axile roots, common QTLs existed for ER(Ax) and L (Ax). Quantitative trait loci for the elongation rates of axile roots responded more clearly to water stress compared to root length. Two major QTLs were detected: a QTL for general vigor in bin 2.02, affecting most of the traits, and a QTL for the constitutive increase in k (Lat) and k (Lat)/ER(Ax) in bins 6.04-6.05. The latter co-located with a major QTL for the anthesis-silking interval (ASI) reported in published field experiments, suggesting an involvement of root morphology in drought tolerance. Rapid seedling tests are feasible for elucidating the genetic response of root growth to low water potential. Some loci may even have pleiotropic effects on yield-related traits under drought stress.

Mapping of QTLs for lateral and axile root growth of tropical maize.

Maize genotypes may adapt to dry environments by avoiding desiccation by means of a deeper root system or by maintaining growth and water extraction at low water potentials. The aim of this study was to determine the quantitative genetic control of root growth and root morphology in a population of 236 recombinant inbred lines (RILs) from the cross between CML444 (high-yielding) x SC-Malawi (low-yielding), which segregates for the response to drought stress at flowering. The RILs and the parental lines were grown on blotting paper in growth pouches until the two-leaf stage under non-stressed conditions; the parents were additionally exposed to desiccation stress induced by polyethylene glycol with a molecular weight of 8000 Dalton (PEG-8000). The lengths of axile and lateral roots were measured non-destructively at 2, 5, 7 and 9 days after germination, by scanning with an A4 scanner followed by digital image analysis. CML444 had a lower rate constant of lateral root elongation (k(Lat)) than SC-Malawi, but the two genotypes did not differ in their response to desiccation. QTLs affecting root vigor, as depicted by increments in k(Lat), the elongation rate of axile roots (ER(Ax)) and the number of axile roots (No(Ax)) were identified in bins 2.04 and 2.05. QTLs for No(Ax) and ER(Ax) collocated with QTLs for yield parameters in bins 1.03-1.04 and 7.03-04. The correspondence of QTLs for axile root traits in bins 1.02-1.03 and 1.08 and QTLs for lateral roots traits in bins 2.04-2.07 in several mapping populations suggests the presence of genes controlling root growth in a wide range of genetic backgrounds.

Drought stress and tropical maize: QTL-by-environment interactions and stability of QTLs across environments for yield components and secondary traits.

A recombinant inbred line (RIL) population was evaluated in seven field experiments representing four environments: water stress at flowering (WS) and well-watered (WW) conditions in Mexico and Zimbabwe. The QTLs were identified for each trait in each individual experiment (single-experiment analysis) as well as per environment, per water regime across locations and across all experiments (joint analyses). For the six target traits (male flowering, anthesis-to-silking interval, grain yield, kernel number, 100-kernel fresh weight and plant height) 81, 57, 51 and 34 QTLs were identified in the four step-wise analyses, respectively. Despite high values of heritability, the phenotypic variance explained by QTLs was reduced, indicating epistatic interactions. About 80, 60 and 6% of the QTLs did not present significant QTL-by-environment interactions (QTL x E) in the joint analyses per environment, per water regime and across all experiments. The expression of QTLs was quite stable across years at a given location and across locations under the same water regime. However, the stability of QTLs decreased drastically when data were combined across water regimes, reflecting a different genetic basis of the target traits in the drought and well-watered trials. Several clusters of QTLs for different traits were identified by the joint analyses of the WW (chromosomes 1 and 8) and WS (chromosomes 1, 3 and 5) treatments and across water regimes (chromosome 1). Those regions are clear targets for future marker-assisted breeding, and our results confirm that the best approach to breeding for drought tolerance includes selection under water stress.

Low temperature stress in maize (Zea mays L.) induces genes involved in photosynthesis and signal transduction as studied by suppression subtractive hybridization.

Unfavourable environmental conditions such as cold induce the transcription of a range of genes in plants in order to acclimate to these growth conditions. To better understand the cold acclimation of maize (Zea mays L.) it is important to identify components of the cold stress response. For this purpose, cold-induced genes were analysed using the PCR-select cDNA subtraction method. We identified several novel genes isolated from maize seedling exposed for 48h to 6 degrees C. Of 18 Zea mays cold-induced genes (ZmCOI genes) characterized, the majority share similarities with proteins with known function in signal transduction and photosynthesis regulation. RT-PCR was conducted for a selected group of genes, namely ZmCOI6.1, ZmACA1, ZmDREB2A and ZmERF3, confirming the induction by low temperature. In addition, it was found that their expression was strongly induced by other abiotic stresses such as drought and high salt concentration, by stress signalling molecules such as jasmonic acid, salicylic acid and abscisic acid, and by membrane rigidification. These results suggest that this group of genes is involved in a general response to abiotic stresses.

The use of green fluorescent protein as a tool to identify roots in mixed plant stands.

Roots take up most of the resources required by a plant, but a lack of efficient research tools hinders our understanding of the function and relevance of the root system. This is especially evident when the research focus is not on a single plant, but on multiple plants that share the same soil resources. None of the available methods allow for simple, inexpensive, non-destructive, and objective assignment of observed roots in a mixture of plants to a target plant. Here, we demonstrate that transgenic plants expressing the green fluorescent protein (GFP), combined with the well established minirhizotron technique, is a route to overcoming this limitation. We planted transgenic maize (Zea mays L.) in combination with either its corresponding wild type, Italian ryegrass (Lolium multiflorum Lam.), or soybean (Glycine max (L.) Merr.). Identification of fluorescent roots allows the relative distribution of roots of each plant type and their interaction and interference with each other to be observed. The selected plants are suitable for model experiments to unravel fundamental belowground ecological processes. Because genetic transformation of plants is an established technique that can be applied to a large set of plant species, this method will be of interest to a broad range of research areas.

QTL studies reveal little relevance of chilling-related seedling traits for yield in maize.

Prolonged low temperature phases and short-term cold spells often occur in spring during the crucial stages of early maize (Zea mays L.) development. The effect of low temperature-induced growth retardation at the seedling stage on final yield is poorly studied. Therefore, the aim was to identify genomic regions associated with morpho-physiological traits at flowering and harvest stage and their relationship to previously identified quantitative trait loci (QTLs) for photosynthesis and morpho-physiological traits from the same plants at seedling stage. Flowering time, plant height and shoot biomass components at harvest were measured in a dent mapping population for cold tolerance studies, which was sown in the Swiss Midlands in early and late spring in two consecutive years. Early-sown plants exhibited chilling stress during seedling stage, whereas late-sown plants grew under favorable conditions. Significant QTLs, which were stable across environments, were found for plant height and for the time of flowering. The QTLs for flowering were frequently co-localized with QTLs for plant height or ear dry weight. The comparison with QTLs detected at seedling stage revealed only few common QTLs. A pleiotropic effect was found on chromosome 3 which revealed that a good photosynthetic performance of the seedling under warm conditions had a beneficial effect on plant height and partially on biomass at harvest. However, a high chilling tolerance of the seedling seemingly had an insignificant or small negative effect on the yield.

Effect of the 1BL.1RS translocation on the wheat endosperm, as revealed by proteomic analysis.

The introduction of the 1RS chromosome of rye into wheat made wheat more resistant to several pathogens. Today, this resistance has been overcome but the 1BL.1RS translocation remains interesting because of the improved yield and despite the lower rheological properties it produces. Nothing has been reported yet on the impact of rye chromatin introgression on the grain proteome of wheat. The comparison of the 2-DE profiles of 16 doubled haploid lines, with or without the 1BL.1RS translocation, revealed quantitative and qualitative proteic variations in prolamins and other endosperm proteins. Eight spots were found specifically in lines having the 1BL.1RS translocation; 16 other spots disappeared from the same lines. Twelve spots, present in both genotypes, met the criteria for up- or down-regulated spots. In translocated genotypes, a highly overexpressed spot, identified as a gamma-gliadin with nine cysteine residues, suggests that the lack of LMW-GS induced by 1BL.1RS is counterbalanced by an overexpression of a relatively similar prolamin. Moreover, a spot that was absent from 1BL.1RS genotypes was identified as a dimeric alpha-amylase inhibitor. It was considered to be a valuable candidate to explain the sticky dough associated with translocated cultivars.

Mapping of quantitative trait loci associated with chilling tolerance in maize (Zea mays L.) seedlings grown under field conditions.

The effect of low growth temperature on morpho-physiological traits of maize was investigated by the means of a QTL analysis in a segregating F(2:3) population grown under field conditions in Switzerland. Chlorophyll fluorescence parameters, leaf greenness, leaf area, shoot dry weight, and shoot nitrogen content were investigated at the seedling stage for two years. Maize was sown on two dates in each year; thus, plants sown early were exposed to low temperature, whereas those sown later developed under more favourable conditions. The main QTLs involved in the functioning of the photosynthetic apparatus at low temperature were stable across the cold environments and were also identified under controlled conditions with suboptimal temperature in a previous study. Based on the QTL analysis, relationships between chlorophyll fluorescence parameters and leaf greenness were moderate. This indicates that the extent and functioning of the photosynthetic machinery may be under different genetic control. The functioning of the photosynthetic apparatus in plants developed at low temperature in the field did not noticeably affect biomass accumulation; since there were no co-locations between QTLs for leaf area and shoot dry weight, biomass accumulation did not seem to be carbon-limited at the seedling stage under cool conditions in the field.